Composition comprising cysteamine for improving lactation in dairy animals

ABSTRACT

The use of cysteamine, a salt thereof or a composition containing cysteamine or a salt thereof for improving lactation of lactating animals.

BACKGROUND OF INVENTION

1. Field of Invention

The present invention relates to the uses of cysteamine or acysteamine-containing composition for improving lactation of lactatinganimals, and in particular, but not limited to, dairy animals such asdairy cows. The present invention also relates to a feed and a methodfor improving lactation of lactating animals.

2. Description of Prior Art

Studies have indicated that growth hormones play an important role inregulating lactation in dairy animals such as diary cows. Furtherstudies on dairy animals have shown that milk production therefrom canbe increased when exogenous growth hormones are administered to theanimals. However, there are a number of disadvantages in directly usinggrowth hormones in increasing milk production. Firstly, growth hormonesfrom different animals are seldom homogenous and different dairy animalsonly react to certain types of specific growth hormones. Since suitableexogenous growth hormones are normally extracted from pituitary glands,it is rather difficult and uneconomical to prepare sufficient quantityof suitable exogenous growth hormones for use on a large-scaleapplication. Although exogenous growth hormones can now be preparedusing DNA recombinant technology, exogenous growth hormones manufacturedby such method are still rather expensive. Secondly, the administrationof exogenous growth hormones into dairy animals is normally performed bydirect injection, which is inevitably rather costly and difficult toadminister in a large farm. Thirdly, it is rather difficult to controlthe dose administered to produce precisely the desired effect, and anoverdose of exogenous growth hormones is likely to be harmful to theanimals. Fourthly, residuals of these exogenous growth hormones may bepassed to the dairy products and subsequently to humans throughconsumption thereof. Further studies in this regard are requiredalthough some scientists are concerned about the negative side effectsof these exogenous growth hormones to humans.

Cysteamine is a component of co-enzyme A and works as a physiologicalregulator. Cysteamine has been used as an additive in feed in promotinggrowth of meat-producing animals. U.S. Pat. No. 4,711,897 disclosesanimal feed methods and feed compositions comprising cysteamine.

However, it has been identified that cysteamine is a fairly sensitiveand unstable compound under normal room temperature conditions. Forexample, cysteamine is readily oxidized when exposed to air or at anelevated temperature. Cysteamine is highly hydroscopic. Also, cysteamineis unpalatable when taken directly by mouth. Further, ingestingcysteamine directly will cause undesirable gastro side effects. Forthese reasons, the use of cysteamine had for a long time been limited todirect injection of cysteamine-containing solution into meat-producinganimals.

PRC Patent Publication No. CN1358499 and International Publication No.WO/0248110 disclose an improvement of a cysteamine-containingcomposition which can be mixed with basal animal feed to promote growthand increase in animal weight. However, there continues to exist a needfor a composition, feed and/or method for increasing lactation by dairymammals and particularly dairy cows. Preferably, the composition andmethod are safe and can be easily administered and inexpensive to carryout.

It is thus an object of the present invention in which the above issuesare addressed, or at least to provide a useful alternative to thepublic.

SUMMARY OF INVENTION

According to a first aspect of the present invention, there is providedthe use of cysteamine, a salt thereof or a composition containingcysteamine or a salt thereof for improving lactation of lactatinganimals.

According to a second aspect of the present invention, there is provideda feed for improving lactation of lactating animals comprising acomposition containing cysteamine or a salt thereof and a stabilizer.

According to a third aspect of the present invention, there is provideda method for improving lactation of lactating animals comprising stepsof producing a final feed by mixing a composition containing cysteamineor a salt thereof and a stabilizer with a suitable basal feed for theanimals, and feeding the animals with the final feed.

The improving lactation may be an increase in milk yields, fat-correctedmilk yields, milk fat content therein and/or milk protein contenttherein.

Preferably, the composition comprises substantially 1 to 95 wt %cysteamine having the chemical formula of NH₂—CH₂—CH₂—SH, or a saltthereof. In particular, the composition may comprise substantially 30 wt% cysteamine.

Suitably, the composition comprises substantially 1 to 80 wt % of thestabilizer. The stabilizer is preferably selected from a group includingcyclodextrin or a derivative thereof. In particular, the composition maycomprise substantially 10 wt % of the stabilizer.

Advantageously, the composition further comprises ingredient(s) selectedfrom a group including a bulking agent, a disintegration agent and acoated carrier. Preferably, the coated carrier in some embodiments is asolid carrier which is a coating soluble in intestines of the animals.The coated carrier suitably exhibits a multi-layer structure in thecomposition. The coated carrier is preferably adapted to remainundissolved at pH 1.5 to 3.5.

Preferably, the feed comprises substantially 400 to 1000 ppm of thecomposition, or substantially 1000 to 2174ppm of the composition when inits dry state. Alternatively, the feed comprises substantially 120 to300 ppm cysteamine, or substantially 200 to 650 ppm cysteamine when inits dry state.

In some embodiments, the feed comprises other foodstuffs selected from agroup including normal premix, cornmeal, cotton seed, wheat gluten,maize silage rutabaga, sugar beet pulp, apple pulp, ryegrass, fescuegrass, alfalfa, feed concentrate and feed supplement.

In one embodiment, the mixing comprises directly mixing the compositionwith the basal feed. In another embodiment, the mixing comprises firstlypreparing a premix including cysteamine or the composition, andsubsequently mixing the pre-mix with the basal feed. The premix isprepared by mixing cysteamine or the composition with a food materialsuch as cornmeal. The premix preferably comprises 5 to 25 wt % of thecomposition. In particular, the premix may comprise 10 to 20 wt % of thecomposition.

Preferably, the animals are fed with substantially 5.64 to 12.71 gcysteamine, or a salt thereof, or 19.79 to 42.36 g of the composition,per animal per day. Alternatively, the animals are fed with twice theamount of cysteamine or the composition but the frequency of theadministration thereof can be reduced to every other day instead ofevery day. Similar results would be produced.

The lactating animals referred above may be dairy cows.

BRIEF DESCRIPTION OF DRAWING

The invention will now be described, by way of non-limiting examplesonly, with reference to the accompanying drawing, in which:

FIG. 1 is a graph showing level of lactation of three groups of dairycows in an experiment.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is based on the demonstration that cysteamine or acysteamine-containing composition when administered to lactating animalssuch as diary cows has activity in improving lactation therefrom. Thelactating animals or dairy animals referred hereinafter include anymilk-producing animals such as dairy cows. Prior to this finding, therewas no suggestion or sufficient indication that cysteamine or itsvariants or derivatives might have such activity. The present inventionalso provides a feed and a method for improving lactation of lactatinganimal. The use of the present invention also prolongs and heightens themilk yield during later stage of lactation of dairy animals. Theinvention may be practiced by directly mixing cysteamine or thecysteamine-containing composition with a suitable basal feed.Alternatively, the invention may be practiced by mixing firstly a premixmade of cysteamine or the cysteamine-containing composition and otheringredients such as cornmeal, and secondly the premix with a suitablebasal feed to form a final feed.

One effect of cysteamine is described in PRC Patent Publication No.CN1358499, the content of which is incorporated herein. However, apreviously unknown effect of cysteamine is its effect on improvinglactation of lactating animals. The effect of cysteamine or acysteamine-containing composition on lactating animals is explained asfollows. It is believed that cysteamine having a physiological activityacts as a growth stimulator. Natural cysteamine is a part of coenzyme A(also know as CoA-SH or CoA) which is a coenzyme pattern of pantothenicacid. In the course of metabolism, coenzyme A acts as the carrier ofdihydrosulfuryl or variants of hydrosulfuryl which is linked with thehydrosulfuryl of coenzyme A. Experiments performed on other animals suchas pigs, poultry, fowls, goats, rabbits and fish have shown thatcysteamine can deplete somatostain (SS) in the animals. This increasesthe level of growth hormone in the blood of the animals which at thesame time raises the level of various other growth stimulating factorsincluding insulin-like growth factor I (IGF-I), insulin,triiodothyronine (T3), trthyroxine (T4) and beta-endorphin (beta-END).As regards the present invention, the growth hormone is believed todirectly stimulate the development and activity of mammalian glands ofdairy animals and the maintenance of lactation therefrom.

With the increase of these various growth-promoting factors, thedigestive metabolic rate of the animals is correspondingly increased. Itis understood that the general protein synthesis rate of the animals isaccordingly increased, and thus lactation is increased.

The cysteamine-containing composition used in the present inventioncomprises two main ingredients of 1 to 95 wt % of cysteamine (or itssalts, for example, cysteamine hydrochloride, or other pharmaceuticallyacceptable acid addition salts thereof) and 1 to 80 wt % of a carriersuch as an inclusion compound host material. The chemical formula ofcysteamine is HSCH₂CH₂NH₂. The term “cysteamine” referred hereinaftermeans cysteamine and/or its salt like compounds. Cysteamine and itssalts are well known in the chemical literature.

The general chemical formula of a cysteamine salt is C₂H₇NS.X, where Xmay be HCl, H₃PO₄, bitartrate, salicylate, etc. The cysteamine used ispreferably of pharmaceutically acceptable standard and the content ofcarbon, hydrogen, nitrogen and sulfur therein are substantially 31.14 wt%, 9.15 wt %, 18.16 wt % and 41.56 wt % respectively. While the workablecontent of cysteamine in the cysteamine-containing composition rangesfrom 1 to 95 wt %, a preferable range of 1 to 75 wt % and a morepreferable range of 1 to 40 wt % of cysteamine may be used. Cysteamineis one of the main active ingredients of the cysteamine-containingcomposition. However, it has been identified that if the content ofcysteamine in the cysteamine-containing composition exceeds 95 wt %,mixing the composition with a basal feed would be rather difficult andthe effect of the composition for regulating lactation of the dairyanimals would be hindered.

The inclusion compound host material may comprise mainly cyclodextrinand/or its derivatives which are selected from a group including methylβ-cyclodextrin (M-β-CD), hydropropyl β-cyclodextrin (HP-β-CD),hydroethyl β-cyclodextrin (HE-β-CD), polycyclodextrin, ethylβ-cyclodextrin (E-β-CD) and branched cyclodextrin. The general chemicalformula of cyclodextrin is (C₆O₅H₉)_(n).(C₆O₅H₉)₂ and the structuralformula is as follows.

where α-CD n=4; β-CD n=5; γ-CD n=6.

(Cyclodextrin is a cyclic oligomer of alpha-D-glucopyranose.)

It is worthwhile to note that the β-CD form of cyclodextrin ispreferably used because the internal diameter of its molecule is about6-8 Å which makes it a particular suitable candidate as an inclusioncompound host material for preparation of the cysteamine-containingcomposition, which involves the use of an inclusion process. The term“cyclodextrin” referred hereinafter means cyclodextrin and/or itsderivatives. Any derivative of cyclodextrin which has the property ofstabilizing and protecting cysteamine from degradation may be used. Forexample, any one of the group of cyclodextrin or its derivativesmentioned above may be used.

While the workable content of the inclusion compound host materials inthe cysteamine-containing composition ranges from 1 to 80 wt %, apreferable workable range of 1 to 60 wt % and a more preferable workablerange of 10 to 40 wt % of the inclusion compound host materials may bealso be used. The actual amount of the inclusion compound host materialsused will depend on the actual content of the cysteamine used inpreparing the cysteamine-containing composition.

The cysteamine-containing composition may also comprise 1 to 90 wt % offillers although a preferable workable range of 1 to 60 wt % and a morepreferable workable range of 1 to 40 wt % of the fillers may also beused in the composition. The actual content will depend on the actualamount of cysteamine and inclusion compound host materials used. Thefillers may be selected from a group including powdered cellulose,starch and calcium sulfate (e.g. CaSO₄.2H₂O). It is to be noted that ifthe content of the fillers exceeds 90 wt % in the cysteamine-containingcomposition, the content of the main active ingredients will thus bereduced, and the cysteamine-containing composition may becomeineffective in improving lactation of the animals fed with a feed mixedtherewith.

The cysteamine-containing composition may also comprise 5 to 50 wt % ofdisintegrants and binders although a preferable workable range of 10 to40 wt % and a more preferable workable range of 15 to 35 wt % may alsobe used. The actual content will depend on the actual amount ofcysteamine, the inclusion compound host material and other ingredientsused. The binders and disintegrants may be selected from a groupincluding hydropropyl starch, microbial alginate, microcrystallinecellulose and starch. It has been identified that if the content of thedisintegrants and binders in the composition is less than 5 wt %,granules of the composition produced will lack the required hardness. Inaddition, manufacturing of the composition would become very difficult.If however the content of the disintegrants and binders is more than 50wt %, the resulting composition will have excessive hardness, this isespecially so if the content of binders represent a large portion of themixture of the disintegrants and binders. This will result in difficultabsorption of the composition by the intestines of the animals.

The cysteamine-containing composition may also comprise 0.05 to 0.3 wt %of flavoring and smelling agents which may be a flavoring essence.

The cysteamine-containing composition may also comprise 1 to 20 wt % ofcoating materials although a preferable workable range is 1 to 15 wt %and a more preferable workable range is 2 to 10 wt %. The actual contentwill depend on the actual amount of cysteamine, the inclusion compoundhost materials and the other ingredients used. The coating materials arepreferably enteric-coated which allows dissolution in an alkalineenvironment such as in the intestines. The coating materials may be madeof and selected from a group including cellulose acetate phthalate,starch acetate phthalate, methyl cellulose phthalate, glucose orfructose derivatives from phthalic acid, acrylic and methacryliccopolymers, polymethyl vinyl ether, partly esterified substance ofmaleic anhydride copolymers, lac and formogelatine. It has beenidentified that if the content of the coating materials is less than 1wt %, granules of the composition may not be entirely covered by thecoating materials which act as a protective layer. Thecysteamine-containing composition may thus degrade before being absorbedby the intestines into the bloodstream of the animals. On the otherhand, if the content of the coating materials exceeds 15 wt %, theactive ingredients in the composition may not effectively be releasedfrom the composition. Thus, the intended regulation of lactation wouldnot be achieved.

The cysteamine-containing composition used in the present invention isin the form of small granules each of which has a preferable diameter ofsubstantially 0.28 to 0.90 mm. These granules are prepared using amicro-encapsulation method. The method involves using a macromolecularsubstance having inclusion property. One substance which may be used isthe inclusion compound host material (which may comprise mainlycyclodextrin) is described above. The inclusion compound host materialis a macromolecular substance which acts as a molecular capsule toengulf the molecules of cysteamine, whereby cysteamine in thecomposition is protected and insulated from light, heat, air andmoisture of the surroundings. The stability of cysteamine is thuspreserved. The inclusion compound host material used in themicro-encapsulation method is preferably a cyclic polysaccharidecompound having 6 to 12 glucose molecules, which is produced by reactingcyclodextrin glycosidtransferase and starch in the presence of Bacillus.Various studies using acute, sub-acute and chronic toxic tests haveshown that the macromolecular substance is non-toxic. Subsequent to themicro-encapsulation process, each granule may be coated with at leastone and preferably a plurality of layers of the coating materialsdescribed above. The following provides a more detailed description of amethod of preparing the cysteamine-containing composition used in thepresent invention.

In a jacketed reactor linked with polytetrafluoroethylene and equippedwith a polytetrafluoroethylene coated stirrer, 4080 g of 75 wt %cysteamine hydrochloride solution in ethanol is added with mainlynitrogen being the atmosphere. The purity, melting point and burningresidue of the cysteamine used are preferably 98% or above, 66 to 70° C.and 0.05% or below respectively. 1200 g β-cyclodextrin is then addedinto the reactor similarly under the protection of nitrogen gas. (Thequality of β-cyclodextrin is in accordance with the requirements for afood additive. In particular, the dry basis purity is more than 98%; theweight loss by drying is less than 10.0%; the burning residue is lessthan 0.2%; the content of heavy metal is less than 10 ppm; the arseniccontent is less than 2 ppm.) The mixture is then heated for 3 hours at40° C. Heating is then stopped and stirring continues for two hoursthereafter, products resulted therefrom are then grounded and sievedthrough a screen (e.g. 40-mesh) filter after the products have beenvacuum dried at a temperature of 40-50° C. All parts of the equipment,which may come in contact with the ingredients of the composition,should preferably be made of stainless steel.

In a tank-type mixer, 4200 g (on dry basis) of the cysteamine which hasundergone the inclusion process as described, 2600 g of the fillers, and1200 g of the disintegrants and 1700 g binders are added under theprotection of a dry surroundings. These ingredients are then thoroughlymixed, and a suitable amount of anhydrous ethanol may be added and thenmixed therewith. The resulting mixture presents a soft material withmoderate hardness, so that it can be shaped into a ball by a light holdof palms. The ball-shaped resulting mixture may then be broken up by alight touch. After the mixture is pelleted by a granulator under theprotection of nitrogen, the small granules resulting therefrom isimmediately introduced to a fluid-bed dryer, and is then dried at thetemperature of 40-50° C. in a substantially vacuum environment.

Enteric coating materials are then prepared by a method with thefollowing formulation: cellulose acetate phthalate 8.0 g, polyethyleneglycol terephthalate 2.4 ml, ethyl acetate 33.0 ml and isopropyl acetate33.6 ml. The resultant granules obtained above are uniformly coatedunder the protection of nitrogen with at least one layer but preferablya plurality of layers of the enteric coating materials described above.The enteric coating materials are dissolvable only at an alkalineenvironment. This can prevent the cysteamine from prematurely escapedfrom the composition while it is still in the stomach of the animal.Cysteamine can adversely stimulate gastric mucous of the stomach of theanimals.

The resultant granules of the cysteamine-containing composition are thendried completely in a substantially vacuum dryer at a temperature of 40to 50° C. Then, all solvents are removed. The resultant granules arethen allowed to cool to room temperature, the micro-capsula were mixedwith a suitable amount of flavoring and smelling agents by a cantileverdouble helix blender. The cysteamine-containing composition is amicrocapsule with its interior having cysteamine hydrochloride andcyclodextrin, and with its exterior coated with the enteric coatingmaterials.

The composition produced will exhibit small granular (ormicro-particulate) shape having smooth surface, good flow property, andis easy to be blended with various animal feeds. The diameter of eachgranule of the composition is preferably 0.28 to 0.90 mm. Thecomposition also has excellent stability. It has been found that afterthe composition is packaged with sealed plastic bags and stored for oneyear in a cool, dark and dry place, their properties remain unchanged.Therefore, they meet the requirements for a feed additive.

The composition having the particular construction described above has anumber of functional advantages over cysteamine by itself. Firstly, theactivity of the cysteamine contained in the composition is preservedafter it has been produced. This is important as feed additive such asthe composition may be stored for a relatively long period of timebefore use. Secondly, the composition does not cause any noticeablegastro side effects to the animals fed therewith. Thirdly, the activityof the composition is preserved not only during storage but moreimportantly until it reaches the intestines of the animals. Fourthly,the composition can be easily administered to farm animals on a largescale basis cost-effectively because it can be readily mixed with anybasal feed. No separate procedure or injection is needed at all.

Various experiments have been conducted to demonstrate thatadministering a feed having cysteamine or a cysteamine-containingcomposition increases lactation from dairy animals, three experiments ofwhich are described in detail as follows.

EXPERIMENTS Experiment 1

Background Information

The experiment was conducted in a private dairy farm located inGuangming, PRC in April and May 2001. Thirty dairy cows were randomlyselected for the experiment. Before the experiment, the cows weresimilar in weight, age, milk yield, and milk fat content and havesimilar calving number. All the cows had had approximately five monthsof lactation. The cows were randomly divided into three groups of ten,namely Group I, Group II and Group III, Group III being in the control.Prior to the experiment, there was no statistical difference (p>0.05) onaverage milk yield among the three groups of cows. Table 1 below showsdata in respect of the three groups of cows and milk produced therefromprior to the experiment. TABLE 1 Characteristics of diary cows beforeexperiment Group Characteristics Group I Group II Group III Number ofcows 10 10 10 Average number of calving  1  1  1 Average lactationperiod (days) 212.8 ± 5.20  208.4 ± 4.05  208.3 ± 3.91  Average milkyield (kg/day) 27.5 ± 4.27 27.4 ± 4.05 27.5 ± 3.97 FCM (3.5%, kg/day)28.636 ± 5.246  28.773 ± 5.777  28.978 ± 4.973  Average fat content inFCM (wt %) 3.76 ± 0.41 3.78 ± 0.37 3.84 ± 0.39 Average protein contentin milk (g/day) 856.6 ± 109.1 964.7 ± 159.6 869.8 ± 112.7 Average numberof somatic cells in milk 91.7 ± 58.7  86.3 ± 49.99 84.9 ± 58.5(×1000/ml)Note:FCM = fat corrected milk

It is to be noted that during a lactation period, the content of milkproteins and other nutrients in milk may often remain relatively stable,whereas the fat content can vary substantially. In this regard, it hasbecome one of the international standards to express the quantity ofmilk produced in terms of its equivalent quantity of fat corrected milk(FCM) for unified comparison. In the experiment, the quantity of milkproduced by the three groups of dairy cows are expressed both in itsoriginal quantity (see Table 7) as well as quantity in 3.5 wt % FCM (seeTable 8). It is however to be noted that there are other standards forquantifying milk production.

Materials

The recipe of two basal feeds used in the experiment are shown in Table2 below, and the details of the respective diets of the three groups ofcows are described below. TABLE 2 Composition of basal feeds For usebefore Apr. 30, For use on or after Apr. 30, Ingredients (kg) 2001 2001Basal concentrate 5 4.5 Supplement feed 3 3 Normal premix 1.4 1.3 Cottonseed 0.4 0.2 Wheat gluten 10 11 Maize silage 14 14.5 Rutabaga 4 5 Sugarbeet pulp 0.5 0.4 Apple pulp 0.5 0.4 Ryegrass 4 — Fescue grass 3 3Alfalfa 0.8 0.5 Total (Basal feed) 46.6 43.8

Two batches of the basal feeds were prepared, the first batch was usedin the first period of the experiment, i.e. from 11 Apr. to 29 Apr. 2001while the second batch was used in the second period of the experiment,i.e. from 30 Apr. to 29 May 2001. The different basal feed was used inthe second period of the experiment in order to suit the physiologicalrequirements of the dairy cows at that particular developmental stage.As shown in Table 2 above, the first batch of feed comprised a pluralityof ingredients including basal concentrate, feed supplement, normalpremix, cottonseed, wheat gluten, maize silage, rutabaga, sugar beetpulp, apple pulp, ryegrass, fescue grass and alfalfa. The normal premixis formulated to compose ingredients such as vitamins and minerals,which are suitable for the cows in a particular physiological stage. Thebasal concentrate, supplement feed and normal premix were nutritionadditives containing such as vitamins and rare elements, which wereadded in the feed in accordance with America's Animal National Nutritionstandard. The second batch of basal feed generally comprised the sameingredients although the quantities thereof were different.

The basal feeds were used to prepare final feeds. Referring to Table 3below, the final feeds comprised 200 g of a specific premix which wasprepared by mixing a cysteamine-containing composition with a suitablefood material such as cornmeal. It is to be noted that the specificpremix was different from the normal premix in the basal feed. TABLE 3Composition of final feeds for Groups I and II cows For use before Apr.For use on or Ingredients (kg) 30, 2001 after Apr. 30, 2001 Specificpremix 0.2 0.2 Basal feed 46.6 43.8 Total (final feed) 46.8 44.0

There were however three formulas of the specific premix, each preparedwith different amounts of the cysteamine-containing composition and thefood material. These formulas are illustrated as follows.10 wt % cysteamine-containing composition/90 wt % cornmeal  Formula 1:20 wt % cysteamine-containing composition/80 wt % cornmeal  Formula 2:0 wt % cysteamine-containing composition/100 wt % cornmeal  Formula 3:

While formulas 1 to 3 of the specific premix comprises up to 20 wt % ofthe cysteamine-containing composition, studies have shown that, inpractice, the specific premix may have a content of 5 to 25 wt % of thecysteamine-containing composition and a similar effect on increasinglactation of the dairy cows will result as illustrated below.

Tables 4 to 6 below further summarize the respective diets of the GroupsI to III cows during the experiment. Based on the data in Table 4, it iscalculated that the concentrations of the cysteamine-containingcomposition in the final feeds (or diets) of the Group I cows in thefirst and second periods of the experiment were 427 and 455 ppmrespectively. Based on the data in Table 5, it is calculated that theconcentrations of the cysteamine-containing composition in the finalfeeds (or diets) of the Group II cows in the first and second periodswere 855 and 909 ppm respectively. Referring to Table 6, nocysteamine-containing composition was added in preparing the 180 g ofspecific premix (Formula III). Therefore, the final feeds (or diets) ofthe Group III cows did not contain any cysteamine-containingcomposition. TABLE 4 Diets of the Group I dairy cows For Use Before ForUse On Ingredients April 30 or After April 30 Basal feed (g) 46600 43800Specific premix (g) 200 200 Amount of cysteamine- 10 10 containingcomposition (formula I) in specific premix (wt %) Total diet per day (g)46800 44000

TABLE 5 Diets of the Group II dairy cows Ingredients Before April 30 Onor After April 30 Basal feed (g) 46600 43800 Specific premix (g) 200 200Amount of cysteamine- 20 20 containing composition (formula II) inspecific premix (wt %) Total diet per day (g) 46800 44000

TABLE 6 Diets of the Group III (control) dairy cows Ingredients BeforeApril 30 On or After April 30 Basal feed (g) 46600 43800 Specific premix(g) 180 180 Amount of cysteamine- 0 0 containing composition (formulaIII) in specific premix (wt %) Total diet per day (g) 46780 43980

The cysteamine-containing composition being in mini-pill form comprisedabout 30 wt % cysteamine together with other ingredients includingcyclodextrin which served as a stabilizer. The content of cyclodextrinin the composition was 10 wt %. The composition was prepared by WalcomBio-Chemicals Industry Limited.

To form the final feeds, the appropriate formula of the specific premixwas mixed with the suitable basal feed.

Studies have shown that in practice, the cysteamine-containingcomposition may contain 1 to 95 wt % cysteamine. In any event, it ispreferred that the final feed is adjusted to contain approximately 400to 1000 ppm of the composition. Alternatively, the final feed may beadjusted to contain approximately 120 to 300 ppm of cysteamine.

In its dry state, the feed may comprise 1000 to 2147 ppm of thecomposition and 200 to 650 ppm of cysteamine. The composition used inthe experiment comprised 10 wt % cyclodextrin which is mentioned above.However, depending on the actual amount of cysteamine used in preparingthe composition, the composition may contain 1 to 80 wt % cyclodextrin,as well as other ingredients which may include a bulking agent, adisintegration agent and a solid coated carrier. The composition is inthe form of mini-pill having a multi-layer structure. The compositionthus remains relatively stable at room temperature conditions andun-dissolved at a pH as low as 1.5 to 3.5 (such as in a stomachenvironment) after it has been ingested by the animal. However, thecarrier is made of a coating material which is soluble in a higher pHenvironment such as in the intestines.

Procedure

All three groups of cows were kept together under the same conditionswhich included tether feeding and automatic drinking. The cows wereallowed four hours of exercising and feeding every day. The cows werefed three times a day. During each feeding time, the cows were firstlyfed with roughage and then with the respective diets (see Tables 4 to6). The consumption of roughage was not restricted but was monitored.The cows were milked three times a day at 0600, 1400 and 2100.

During the experiment, data of milk yield expressed in its originalamount and in 3.5 wt % FCM, fat content and protein content of the milkas well as the number of somatic cells in the milk were recorded anddetermined.

Results and Discussion

FIG. 1 shows three curves representing 3.5 wt % FCM yield. As clearlyshown in the figure, the Group II cows generally had the highest 3.5 wt% FCM yield among the three groups of cows throughout the experiment.The Group I cows also had a generally higher 3.5 wt % FCM yield ascompared to the Group III cows.

Table 7 below shows the data of average original milk yield of the threegroups of cows. It is calculated that after feeding on the respectivediets including the cysteamine-containing composition, the Groups I andII cows on average increased lactation by 4.86% and 6.88% (p<0.05) whencompared to the Group III cows. Statistically, these are significantincreases. TABLE 7 Comparison of average milk yield (kg/day) PeriodGroup I Group II Group III Before trial  27.5 ± 4.27a  27.4 ± 4.02a 27.5 ± 4.1a 1^(st) to 10^(th) day 28.99 ± 0.43a  29.5 ± 0.60a 27.29 ±0.72b 11^(th) to 20^(th) day 27.98 ± 0.37a  28.9 ± 0.24a 27.17 ± 0.39b21^(st) to 30^(th) day 28.05 ± 0.59a 28.51 ± 0.75a 26.71 ± 0.61b 31^(st)to 40^(th) day 27.23 ± 0.77a 27.48 ± 0.52a 25.85 ± 0.53b 1^(st) to40^(th) day 28.06 ± 0.84a 28.60 ± 0.91a 26.76 ± 0.79b 41^(st) to 49^(th)day (stop 26.09 ± 0.67a 26.12 ± 0.76a 25.42 ± 0.40b feeding cysteamine-containing diet)(Note: In each row, the figures labeled with same letters means thatthere is no significant difference statistically, i.e. p > 0.05; thefigures labeled with different letters means that there is significantdifference statistically, i.e. p < 0.05.)

For the sake of comparison, the original milk yield of the three groupscows shown in Table 7 has been converted to 3.5 wt % FCM yield which isshown in Table 8 below. The following formula is used to convertoriginal milk yield to its corresponding FCM (3.5 wt %) yield.Equivalent FCM=(0.44×daily milk yield)+(16×daily milk yield×fat contentrate) TABLE 8 Comparison of average FCM (3.5% wt) yield (kg/day) GroupIII Period Group I Group II (control) Before trial 28.64 ± 5.25a 28.73 ±5.78a 28.98 ± 4.97a 1^(st) to 10^(th) day 29.51 ± 0.49a 31.07 ± 0.69b28.73 ± 0.78c 11^(th) to 20^(th) day 30.56 ± 0.41a 31.26 ± 0.26a 29.22 ±0.42b 21^(st) to 30^(th) day 30.46 ± 0.67a 30.37 ± 0.81a 28.60 ± 0.65b31^(st) to 40^(th) day 29.23 ± 0.34a 28.84 ± 0.52a 27.52 ± 0.59b 1^(st)to 40^(th) day 29.94 ± 0.63a 30.39 ± 0.98a 28.52 ± 0.68b 41^(st) to49^(th) day(stop 27.10 ± 0.85a 25.91 ± 0.78b 25.82 ± 0.61b feedingcysteamine- containing diet)(Note: In each row, the figures labeled with same letters means thatthere is no significant difference statistically, i.e. p > 0.05; thefigures labeled with different letters means that there is significantdifference statistically, i.e. p < 0.05.)

It is calculated that after feeding on the respective diets includingthe cysteamine-containing composition, the Groups I and II cows had a4.98% (p<0.01) and 6.56% (p<0.01) higher FCM (3.5 wt %) yield whencompared to the Group III cows. Statistically, these are significantincreases.

As illustrated above, the Groups I and II cows significantly improvedtheir lactation in terms of its original milk yield and 3.5 wt % FCMyield after feeding on cysteamine-containing diets.

It is to be noted that after each calving, the lactation of a cownormally lasts for approximately 300 days. The above experimental dataindicates that administering cysteamine/cysteamine-containingcomposition to dairy cows at least heightens lactation. It appears thatcysteamine can also prolong lactation since when the lactation ismaintained at a higher level for a longer time, the lactation should beprolonged to a certain extent, as shown by the curve representing themilk yield of the Group I cows in FIG. 1.

Table 9 below shows the data of average fat content in the milk producedby the three groups of cows. TABLE 9 Comparison of average fat content(wt %) in milk Group III Period Group I Group II (control) Before trial3.76 ± 0.41a 3.78 ± 0.37a 3.84 ± 0.40b 1^(st) to 10^(th) day 3.62 ±0.39a 3.86 ± 0.26b 3.84 ± 0.26b 11^(th) to 20^(th) day 4.10 ± 0.42a 4.00± 0.24a 3.99 ± 0.23a 21^(st) to 30^(th) day 4.06 ± 0.42a 3.90 ± 0.21b3.95 ± 0.16b 31^(st) to 40^(th) day 4.02 ± 0.45a 3.80 ± 0.22b 3.91 ±0.20c 1^(st) to 40^(th) day 3.95 ± 0.45a 3.89 ± 0.23a 3.92 ± 0.21a41^(st) to 49^(th) day (stop 3.69 ± 0.53a 3.53 ± 0.20a 3.68 ± 0.24afeeding cysteamine- containing diet)(Note: In each row, the figures labeled with same letters means thatthere is no significant difference statistically, i.e. p > 0.05; thefigures labeled with different letters means that there is significantdifference statistically, i.e. p < 0.05.)

Before the experiment, there is no significant difference of milk fatcontent among the three groups of cows (p>0.05, see Table 1). It iscalculated that after feeding on the respective diets including thecysteamine-containing composition, the milk produced by the Group I cowshad an average of 0.77% (p<0.05) higher fat content than that of theGroup III cows. It is also calculated that after feeding on therespective diets including the cysteamine-containing composition, themilk produced by the Group II cows had an average of 0.77% (p<0.5) lowerfat content than that of the Group III cows during the experiment,although the difference is not significant statistically. As illustratedabove, there is no significant difference in fat content in milkproduced by the Groups I and II cows after their diets were added withcysteamine/cysteamine-containing composition. This indicates that thequality of milk produced by cows feeding on a cysteamine-containing dietcan at least be maintained if not increased.

It is however to be noted that during the first ten-day period of theexperiment, the Group I cows appeared to have reacted more slowly to itscysteamine-containing diet in terms of the fat content in its milk incomparison to the Group II cows. However, the fat content in the milkproduced by the Group I cows had risen rapidly and peaked at 4.10 wt %during the period of 11^(st) to 20^(th) day of the experiment. To thecontrary, the Group II cows appeared to have reacted relatively morerapidly to its cysteamine-containing diet in the first ten-day period.In particular, the fat content was 3.78 wt % before the experiment butrose to 3.86 wt % in the first ten-day period of the experiment, and thefat content peaked at 4.00 wt %. It is to be noted that the milkproduced by the Group II cows had the lowest fat content among the threegroups when all the cows were stopped feeding on the respectivecysteamine-containing diets. It is thus indicated in the experiment thatcysteamine or a cysteamine-containing composition when administered atan appropriate dose and/or a particular lactation stage can increase thecontent of milk fat in the milk.

Table 10 below shows the data of average milk protein content in milkproduced by the three groups of cows. TABLE 10 Comparison of averagemilk protein content (g/cow/day) Group III Period Group I Group II(control) Before trial 856.6 ± 109a 864.7 ± 159.6a 869.8 ± 112.7a 1^(st)to 10^(th) day 926.9 ± 89.4a 941.8 ± 109.2a 904.7 ± 116.3b 11^(th) to20^(th) day 911.9 ± 80.1a 917.6 ± 126.8a 875.8 ± 82.7b 21^(st) to30^(th) day 928.3 ± 75.8a 903.4 ± 120.7b 878.2 ± 106.9c 31^(st) to40^(th) day 912.8 ± 69.5a 868.7 ± 121.3b 875.2 ± 103.2b 1^(st) to40^(th) day 920.1 ± 28.8a 907.9 ± 30.6a 883.5 ± 24.2b 41^(st) to 49^(th)day (stop 877.1 ± 79.4a 830.9 ± 116.5b 850.7 ± 89.0c feeding cysteamine-containing diet)(Note: In each row, the figures labeled with same letters means thatthere is no significant difference statistically, i.e. p > 0.05; thefigures labeled with different letters means that there is significantdifference statistically, i.e. p < 0.05.)

Before the experiment, the protein content in milk (g/day/cow) producedby the Group III cows was about 869.8 g which was slightly higher thanthat of the Groups I and II cows, although the difference was notsignificant statistically. After the Groups I and II cows were fed withthe respective cysteamine-containing diets, the protein content in theirmilk increased significantly by 4.14% and 2.76% respectively (p<0.05) ascompared to the Group III cows. After the Groups I and II cows stoppedfeeding on the respective cysteamine-containing diets, the proteincontent reduced close to the pre-experiment levels. It is thereforeillustrated that feeding on cysteamine-containing diet not onlyincreases original milk yield, FCM yield and milk fat content but alsomilk protein content.

Table 11 below shows the data of the number of somatic cells in the milkproduced by the three groups of cows. TABLE 11 Comparison of somaticcells (×10³/ml) in milk Period Group I Group II Group III (control)Before trial  91.7 ± 64.9a  86.3 ± 55.3a  84.9 ± 64.7a 1^(st) to 10^(th)day  96.6 ± 92.8a 102.0 ± 78.0a 116.9 ± 129.0a 11^(th) to 20^(th) day 90.0 ± 82.9a  87.2 ± 64.9a  59.9 ± 33.0b 21^(st) to 30^(th) day 122.8 ±113.3a  89.8 ± 60.9b  69.5 ± 42.7b 31^(st) to 40^(th) day 155.2 ± 155.1a 92.1 ± 58.4b  79.0 ± 59.7b 1^(st) to 40^(th) day 116.1 ± 113.0a  92.8 ±63.6b  81.3 ± 76.3b 41^(st) to 49^(th) day 115.2 ± 115.9a  96.2 ± 54.5b 93.9 ± 45.5b(Note:In each row, the figures labeled with same letters means that there isno significant difference statistically, i.e. p > 0.05; the figureslabeled with different letters means that there is significantdifference statistically, i.e. p < 0.05.)

After feeding on the respective cysteamine-containing diets, the milkproduced by the Groups I and II cows had a higher number of somaticcells. However, the difference remained at an acceptable safety level.

In the experiment, Formula II of the specific premix comprised twice asmuch the cysteamine-containing composition. In other words, the Group IIcows were fed with twice as much cysteamine/cysteamine-containingcompound as compared to the Group I cows. While the milk yield didincrease as compared to that of the Group III cows, the increase was nottwice as much as compared to the Group I cows. It is believed there is acertain physiological limit a cow can react to cysteamine. In the eventthat a relatively high amount of cysteamine were taken by a cow, thephysiology would still not react exceeding its limit. This is desirableas the dosage of the cysteamine-containing composition in the diet isnot critical in that an overdose thereof is not dangerous to the animal.Thus, administering the cysteamine-containing composition in dairy cowsis safe and easy to carry out.

It is to be noted that a final feed has a relatively high water content(e.g. 10 to 60 wt % water content) and the precise concentrations ofcysteamine and cysteamine-containing composition may thus vary dependingon the water content thereof and the humidity of the surroundings. Inthis regard, it is worthwhile to express a workable range of theconcentration of cysteamine and cysteamine-containing composition in thefinal feed based on its dry weight (state). For example, studies havedetermined that the concentration of 1000 to 2174 ppm of thecysteamine-containing composition used in the above examples in thefinal feed based on its dry weight will produce the similar results inincreasing lactation as shown in the examples. Similarly, it iscalculated that the concentration of 200 to 650 ppm of cysteamine in thefinal feed based on its dry weight will likewise produce similarresults.

It is also to be noted that while the administration of cysteamine andthe cysteamine-containing composition to the diary cows are performedvia its diet, it is envisaged that cysteamine and thecysteamine-containing composition may Is likewise be fed to the dairycows in the form of pills, tablets or other suitable state separately ortogether with a food material and this would produce the same effect.Studies have shown that the effective amount of cysteamine administeredto raise the dairy cows may be substantially 5.64 to 12.71 g per cow perday. The effective amount of cysteamine-containing composition may besubstantially 18.79 to 42.36 g per cow per day.

Experiment 2

Background Information

100 black & white dairy cows were used in the experiment. Before theexperiment, the average body weight of the dairy cows was about 600 kg;the average lactation period of the cows was about 135^(th) day and theaverage milk production was about 35 kg. The quality of the milkproduced by the cows prior to the experiment was similar. The cows werenot administered with any cysteamine products beforehand.

The cows were equally divided into one test group and one respectivecontrol group. Each of the test and control groups are further dividedinto four sub-groups during their 19^(th) week of lactation, namelyGroups I to IV, according to their actual milk production, calvinghistory and stage of lactation. The prior milk production (MP) of theGroups I to IV cows were MP≦30 kg, 30 kg<MP≦35 kg, 35 kg<MP≦40 kg, 40kg<MP respectively.

Procedure and Materials

The experiment was carried out during the 20^(th) to 32^(nd) week oflactation of the cows. The actual experiment was preceded by a two-dayadaptation period. At different periods during the experiment, the testsub-groups of cows were administered with different amounts of acysteamine-containing composition via a basal feed. The composition isidentical to that used in Experiment 1. In particular, the compositioncomprises substantially 30% wt cysteamine.

The control group of cows was fed with the same basal feed as the testgroup of cows. The amount of basal feed used was adjusted according tothe actual milk production at the time.

All the cows were fed three times daily. During each feeding time, thecows were fed firstly with roughage and then the basal feed mixed withor without the composition. The cows were milked three times daily at0730, 1430 and 2130.

The milk collected was measured for its quantity and analyzed for itsmilk fat content and milk protein content.

Results and Discussion

Tables 12 to 16 below summarize the experimental data of Experiment 2.TABLE 12 Content of milk fat (wt %) produced by the test and controlgroups of cows CCC 20 30 40 60 mean Sub- Group group T C T C T C T C T CI 3.67 ± 0.70 3.77 ± 0.65 3.66 ± 0.79 3.91 ± 0.68 3.63 ± 3.82 ± 0.573.52 ± 0.43 3.45 ± 0.77 3.69 ± 0.09* 3.73 ± 0.20 0.72 II 3.95 ± 0.413.81 ± 0.49 3.87 ± 0.45 3.74 ± 0.60 3.98 ± 3.78 ± 0.47 3.62 ± 0.45 3.45± 0.77 3.89 ± 0.15* 3.73 ± 0.16 0.43* III 3.58 ± 0.38 3.62 ± 0.58 3.51 ±0.56 3.54 ± 0.66 3.73 ± 3.86 ± 0.53 3.65 ± 0.47 3.64 ± 0.33 3.64 ± 0.153.70 ± 0.17 0.46 IV 3.95 ± 0.38^(#) 3.62 ± 0.37 3.82 ± 0.38* 3.54 ± 0.463.92 ± 3.48 ± 0.55 3.89 ± 0.43* 3.37 ± 0.41 3.89 ± 0.12** 3.50 ± 0.120.48** mean mean mean mean mean 3.80 ± 0.49 3.72 ± 0.51 3.73 ± 0.57 3.67± 0.61 3.84 ± 3.75 ± 0.53 3.65 ± 0.45^(#) 3.50 ± 0.47 3.77 ± 0.11 3.66 ±0.12 0.53(Note: CCC = Cysteamine-containing composition, g per cow per day(g/c/d); T = Test group; C = Control group; *p < 0.05; **p < 0.01;^(#)0.05 < p < 0.15)

Referring to Table 12, it is shown that after administered with a feedcontaining the cysteamine-containing composition during the experiment,there was an average increase of milk fat content in the milk producedby the test cows by about 3.01% (=[3.77 wt %−3.66 wt %]/3.66 wt %×100%).In the test Group II of cows, the average increase of milk fat contentwas about 4.29% (=[3.89 wt %−3.73 wt %]/3.73 wt %×100%). In the testGroup IV of cows, the average increase of milk fat content was moresignificant by about 11.14% (=[3.89 wt %−3.50 wt %]/3.50 wt %×100%).Since the Group IV of cows were the cows with the highest milk yieldbefore the experiment, it is shown that the effect of cysteamine on milkfat content is more prominent on the dairy animals with already highermilk production.

Referring to the four test sub-groups of cows administered with 20g/c/d, 30 g/c/d, 40 g/c/d and 60 g/c/d of the composition respectively,it is calculated that the increase in milk fat content in the milkproduced are 2.15%, 1.63%, 2.40% and 4.29% respectively. TABLE 13Content of milk protein (wt %) produced by the test and control groupsof cows CCC 20 30 40 60 mean Sub- Group group T C T C T C T C T C I 3.10± 3.41 ± 0.33 3.06 ± 3.32 ± 0.36 2.96 ± 0.17* 3.22 ± 0.35 2.99 ± 0.152.73 ± 0.81 3.01 ± 0.06* 3.21 ± 0.05 2.33^(#) II 3.12 ± 2.91 ± 0.22 3.14± 2.94 ± 0.24 3.00 ± 0.25^(#) 2.93 ± 0.21 3.00 ± 0.21 3.01 ± 0.22 3.06 ±0.08** 2.94 ± 0.04 0.28* 0.21** III 2.96 ± 2.94 ± 0.25 3.01 ± 2.98 ±0.34 2.91 ± 0.25 2.88 ± 0.27 2.92 ± 0.21 2.89 ± 0.22 2.95 ± 0.06 2.92 ±0.05 0.28 0.34 IV 2.84 ± 2.63 ± 0.13 2.92 ± 2.67 ± 0.16 2.86 ± 0.26**2.66 ± 0.19 2.88 ± 0.27 2.76 ± 0.21 2.88 ± 0.05** 2.67 ± 0.06 0.27*0.26** mean mean mean mean mean 3.03 ± 2.92 ± 0.03 3.00 ± 2.92 ± 0.312.92 ± 0.24 2.90 ± 0.28 2.96 ± 0.21^(#) 2.90 ± 0.33 3.06 ± 0.17* 2.92 ±0.04 2.08* 0.29^(#)(Note: CCC = Cysteamine-containing composition, g per cow per day(g/c/d); T = Test group; C = Control group; *p < 0.05; **p < 0.01;^(#)0.05 < p < 0.15)

Table 13 shows that the average increase of the milk protein content inmilk produced by the test cows was about 4.79% (=[3.06 wt %−2.92 wt%]/2.92 wt %×100%). In particular, the increase of the milk proteincontent in milk produced by the test Group II of cows was about 4.08%(=[3.06 wt %−2.94 wt %]/2.94 wt %×100%). The increase of the milkprotein content in milk produced by the test Group IV of cows was moresignificant by about 7.87% (=[2.88 wt %−2.67 wt %]/2.67 wt %×100%).Since the Group IV cows were the cows with the highest yield of milkproduction before the experiment, it is shown that the effect ofcysteamine on milk protein content is more prominent on the dairyanimals with already higher milk production.

Referring to the dosage effects of the composition on milk proteincontent, the dose of 20 g/c/d increased the milk protein contentsignificantly in the test sub-groups of cows by about 3.77% (=[3.03 wt%−2.92 wt %]/2.92 wt %×100%). TABLE 14 Production of 3.5 wt % FCM milkby the test and control groups of cows CCC 20 30 40 60 mean GroupSubgroup T C T C T C T C T C I 29.4 ± 3.4 28.3 ± 3.7 28.9 ± 4.4 25.4 ±5.0 25.9 ± 4.5 22.4 ± 6.2 22.6 ± 3.3 19.9 ± 7.0 26.7 ± 3.2 24.0 ± 5.2 II34.9 ± 3.4 35.2 ± 3.1 33.4 ± 4.7 33.0 ± 4.0 30.9 ± 5.0 29.9 ± 4.6 26.6 ±5.8 26.0 ± 5.2 31.0 ± 3.2 31.4 ± 3.4 III 37.8 ± 3.1 37.8 ± 4.6 35.5 ±4.3 35.5 ± 5.9 33.7 ± 5.6 34.1 ± 5.9 29.7 ± 5.5 29.4 ± 6.1 34.2 ± 4.034.2 ± 4.7 IV 45.0 ± 4.8 43.6 ± 4.6 42.1 ± 2.0 40.5 ± 6.5 38.7 ± 3.4^(#)34.2 ± 7.3 35.0 ± 2.0^(#) 30.9 ± 5.9 40.2 ± 12.0 37.3 ± 5.8 mean meanMean mean mean 36.1 ± 36.7 ± 5.6 34.4 ± 5.8 34.2 ± 6.4 31.8 ± 6.2 31.0 ±6.6 27.9 ± 6.2 27.1 ± 6.4 32.4 ± 5.6 32.3 ± 5.7 25.8(Note: CCC = Cysteamine-containing composition, g per cow per day(g/c/d); T = Test group; C = Control group; ^(#)0.05 < p < 0.15)

Table 14 shows that the test Groups I and IV of cows increased their 3.5wt % FCM milk yield very significantly by 11.25% (=[26.7 kg/day−24.0kg/day]/24.0 kg/day×100%) and 7.7% (=[40.2 kg/day−37.3 kg/day]/37.3kg/day×100%) respectively, while in the test Groups II and III of cows,the production of 3.5 wt % FCM was not altered significantly as comparedto that of the respective control groups. Since the test Group I of cowswere the cows with the lowest milk yield before the experiment, it isshown that the positive effect of cysteamine on 3.5% FCM yield is moreprominent on dairy animals with lower FCM yield. TABLE 15 Production oforiginal milk (kg/day) by the test and control groups of cows CCC 20 3040 60 Mean Group Subgroup T C T C T C T C T C I 28.4 ± 25.8 ± 1.3 28.1 ±0.6* 23.7 ± 1.3 25.2 ± 0.8^(#) 21.1 ± 1.9 22.0 ± 0.8* 19.7 ± 2.1 25.9 ±0.7** 23.1 ± 0.9 0.5^(#) II 32.2 ± 33.6 ± 0.5 31.2 ± 0.7 31.7 ± 0.5 28.6± 0.8 28.6 ± 0.8 25.9 ± 1.0 26.0 ± 0.9 29.6 ± 0.5 29.9 ± 0.6 0.6^(#) III37.2 ± 36.8 ± 1.0 35.4 ± 0.7 35.2 ± 1.2 32.4 ± 1.1 32.2 ± 1.4 28.9 ± 1.228.6 ± 1.4 33.5 ± 0.7 33.3 ± 0.7 0.6 IV 42.0 ± 43.0 ± 0.7 40.1 ± 1.040.2 ± 1.9 36.3 ± 1.3 34.0 ± 1.9 33.1 ± 1.2 31.3 ± 1.6 37.9 ± 1.0^(#)36.8 ± 1.4 0.7 mean Mean Mean Mean Mean 34.2 ± 35.2 ± 0.8 33.0 ± 0.733.7 ± 0.8 30.1 ± 0.7 29.6 ± 0.8 27.1 ± 0.7 26.9 ± 0.8 31.1 ± 0.3 31.0 ±0.4 0.7(Note: CCC = Cysteamine-containing composition, g per cow per day(g/c/d); T = Test group; C = Control group; *p < 0.05; **p < 0.01;^(#)0.05 < p < 0.15)

TABLE 16 Production of milk protein (kg/day) by the test and controlgroups of cows CCC 20 30 40 60 mean Sub- Group group T C T C T C T C T CI 0.88 ± 0.08 0.88 ± 0.17 0.86 ± 0.09 0.79 ± 0.16 0.74 ± 0.06 0.69 ±0.19 0.67 ± 0.06 0.56 ± 0.27 0.79 ± 0.12^(#) 0.71 ± 0.18 II 1.00 ± 0.110.97 ± 0.10 0.93 ± 0.25 0.93 ± 0.10 0.86 ± 0.13 0.83 ± 0.10 0.77 ± 0.130.78 ± 0.12 0.89 ± 0.12 0.88 ± 0.08 III 1.10 ± 0.12 1.08 ± 0.14 1.06 ±0.14 1.05 ± 0.16 0.94 ± 0.14 0.93 ± 0.17 0.84 ± 0.13 0.83 ± 0.17 0.99 ±0.11 0.97 ± 0.14 IV 1.19 ± 0.09^(#) 1.13 ± 0.07 1.17 ± 0.08^(#) 1.07 ±0.12 1.04 ± 0.09* 0.90 ± 0.10 0.95 ± 0.07^(#) 0.86 ± 0.10 1.09 ± 0.110.99 ± 0.14 mean Mean Mean Mean Mean 1.03 ± 0.15 1.02 ± 0.14 1.01 ± 0.150.98 ± 0.15 0.89 ± 0.15 0.85 ± 0.15 0.78 ± 0.17 0.80 ± 0.14 0.93 ± 0.150.91 ± 0.14(Note: CCC = Cysteamine-containing composition, g per cow per day(g/c/d); T = Test group; C = Control group; ^(#)0. 05 < p < 0.15)

Table 15 shows that the original milk yields of the test Group I of cowsincreased very significantly by 12.12% (=[25.9 kg/day−23.1 kg/day]/23.1kg/day×100%).

Table 16 shows that the actual quantity of milk protein produced by thetest Group I of cows increased by 11.27% (=[0.79 kg/day−0.71kg/day]/0.71 kg/day×100%). However, the data in Tables 12 to 14 is morerepresentative in illustrating the positive effects of cysteamine onimproving lactation of dairy animals. TABLE 17 Effect of the compositionon feed to milk ratio CCC 20 30 40 60 mean Sub- Group group T C T C T CT C T C I 0.52 ± 0.56 ± 0.03 0.54 ± 0.09 0.60 ± 0.08 0.54 ± 0.06* 0.68 ±0.10 0.58 ± 0.08 0.68 ± 0.14 0.54 ± 0.06** 0.63 ± 0.11 0.01 II 0.46 ±0.44 ± 0.03 0.47 ± 0.04 0.46 ± 0.04 0.50 ± 0.05 0.50 ± 0.06 0.51 ± 0.080.50 ± 0.09 0.48 ± 0.06 0.48 ± 0.06 0.04* III 0.39 ± 0.40 ± 0.03 0.41 ±0.03 0.42 ± 0.05 0.42 ± 0.06 0.43 ± 0.06 0.46 ± 0.08 0.47 ± 0.10 0.41 ±0.04 0.43 ± 0.05 0.02 IV 0.35 ± 0.34 ± 0.02 0.36 ± 0.03 0.37 ± 0.05 0.37± 0.03 0.40 ± 0.06 0.40 ± 0.04 0.42 ± 0.05 0.38 ± 0.02 0.39 ± 0.04 0.02mean mean mean mean mean 0.45 ± 0.43 ± 0.06 0.45 ± 0.08 0.45 ± 0.08 0.47± 0.07 0.49 ± 0.10 0.49 ± 0.09 0.50 ± 0.12 0.46 ± 0.07 0.47 ± 0.09 0.07(Note: CCC = Cysteamine-containing composition, g per cow per day(g/c/d); T = Test group; C = Control group)

Table 17 above shows that the feed to milk ratio is generally notaffected whether or not the cows are administered with the composition.This is important from a cost point of view.

Experiment 3

Background Information

Thirty-two black & white dairy cows were used in the experiment and wererandomly divided into four groups, namely test Groups I to III, and acontrol group. Each group had eight cows. The characteristics of milkproduced by the cows before the experiment is summarized in Table 17below. TABLE 18 Dairy performance of the four groups of cows before theexperiment Group Characteristics I II III Control Number of cows  8  8 8  8 Calving number  1  1  1  1 Day of lactation 20 20 20 20 Daily milkproduction,  26.90 ± 1.91  26.84 ± 4.51  26.76 ± 3.77  26.74 ± 4.47kg/cow/day 3.5 wt % FCM, kg/cow/day  28.22 ± 2.37  29.81 ± 2.48  28.35 ±3.99  29.73 ± 3.35 Milk fat content, wt %  3.90 ± 0.56  4.19 ± 0.58 3.87 ± 0.93  4.20 ± 0.78 Daily milk protein 698.88 ± 63.64 754.59 ±49.45 744.93 ± 52.86 713.29 ± 76.97 production, g/cow/day Number ofsomatic cells in 1097.4 ± 1070.2 803.00 ± 1113.4 126.90 ± 87.10  99.90 ±136.50 milk, 10³/mlProcedure and Materials

The cysteamine-containing composition used was the same composition inExperiments 1 and 2 comprising substantially 30 wt % cysteamine.

There were three stages in the experiment. In the first stage of theexperiment, the feed for the test Groups I to III of cows was mixed withthe composition such that each cow in the three test groups was fed with20 g, 30 g and 40 g of the composition respectively daily. Nocysteamine-containing composition was mixed in the feed for the controlgroup of cows. The first stage of the experiment lasted for thirty-sevendays and was preceded by a three-day adaptation period.

During the second stage of the experiment, the composition was not addedto the feed for the three test groups of cows. However, each cow in thecontrol group was fed with 20 g of the composition daily via its feed.The second stage of the experiment lasted for seven days.

During the third stage of the experiment, the three test groups of cowswere fed with the feed mixed with the composition such that 20 g of thecomposition was taken by each cow daily. Each cow in the control groupwas fed with 40 g of the composition every other day via the feed. Thethird stage of the experiment lasted for seven days.

The daily milk yield, milk fat content, milk protein content and somaticcells in milk were measured and recorded. The cows were fed three timesa day and milked also three times a day.

Results and Discussion

Tables 19 to 24 summarize the data from the experiment. TABLE 19 Effectof the composition on original milk yield (kg/cow/day) Group Time TestGroup I Test Group II Test Group III Control Group Before experiment26.90 ± 1.91^(a) 26.84 ± 4.51^(ab) 26.76 ± 3.77^(abc) 26.74 ±4.47^(abcd) Stage one 1^(st) to 10^(th) day 27.52 ± 0.52^(a) 28.05 ±0.20^(b) 27.40 ± 0.56^(ac) 27.89 ± 0.44^(abcd) 11^(th) to 20^(th) day28.87 ± 0.34^(a) 26.39 ± 0.91^(B) 26.59 ± 0.26^(bC) 27.61 ± 0.59^(bd)21th to 30^(th) day 28.45 ± 0.53^(a) 26.77 ± 0.92^(B) 26.57 ± 0.23^(bc)26.49 ± 0.20^(bcD) 31st to 37^(th) day 27.64 ± 0.84^(a) 27.24 ±0.42^(ab) 26.09 ± 0.20^(ac) 27.08 ± 1.12^(abcd) Average: 1^(st) to 28.14± 0.74^(A) 27.15 ± 0.91^(B) 26.76 ± 0.58^(BC) 27.27 ± 0.76^(BCD) 37^(th)day Stage two 26.95 ± 0.52^(a) 25.94 ± 1.07^(ab) 25.44 ± 0.13^(bC) 25.09± 0.64^(bcd) Stage three 25.51 ± 0.32^(a) 27.00 ± 0.42^(B)(Note:In the table, same letters in a row means that the difference of theirvalues is insignificant, i.e. p > 0.05, different letters in a same rowmeans that the difference of their values is significant p < 0.05;different letter size in a column means their values are significant,i.e. p < 0.01)

Table 19 above shows that the test Group I of cows increased theiroriginal milk yield by 7.41% (=[26.95−25.09]/25.09×100%) when comparedto that of the cows in the control group. In particular, the increase inmilk yield was more significant after the first ten-day period. Forinstance, the increase in milk yield of the test Group I of cows duringthe 11^(th) to 20^(th) day and 21^(st) to 30^(th) day was 4.56%(=[28.87−27.61]/27.61×100%) and 7.40% (=[28.45−26.49]/26.49×100%)respectively. It is thus illustrated that administering an appropriatedose of the composition to dairy animals is effective in increasingtheir original milk yield. In particular, the increase is morenoticeable after an initial short period of the administration of thecomposition. TABLE 20 Effect of the composition on quantity of milk fat(kg/cow/day) in milk Group Control Time Test Group I Test Group II TestGroup III Group Before the 3.90 ± 0.56^(a) 4.19 ± 0.58^(ab) 3.87 ±0.93^(abc) 4.20 ± experiment 0.78^(abcd) Stage one 1^(st) to 20^(th) day3.63 ± 0.68^(a) 3.63 ± 0.61^(ab) 4.07 ± 0.54^(abc) 3.63 ± 0.44^(abd)21^(st) to 37^(th) day 4.26 ± 0.59^(a) 4.35 ± 0.78^(ab) 4.36 ±0.76^(abc) 4.17 ± 0.54^(abcd) Average: 1^(st) to 3.95 ± 0.70^(a) 3.99 ±0.77^(ab) 4.21 ± 0.65^(abc) 3.90 ± 37^(th) day 0.55^(abcd) Stage two3.69 ± 0.53^(a) 3.69 ± 0.30^(ab) 3.73 ± 0.82^(abc) 3.58 ± 0.35^(abcd)Stage three 3.94 ± 0.31^(a) 3.98 ± 0.48^(ab78)(Note:In the table, same letters in a row means that the difference of theirvalues is insignificant, i.e. p > 0.05, different letters in a same rowmeans that the difference of their values is significant p < 0.05;different letter size in a column means their values are significant,i.e. p < 0.01)

Table 20 above shows that the milk fat in milk produced by the testGroup III of cows increased by 7.95% (=[4.21−3.90]/3.90×100%) in stageone of the experiment. In particular, the increase in milk fat wasparticularly significant during the first twenty-day period of theexperiment, i.e. 12.12% (=[4.07−3.63]/3.63×100%).

In stage two of the experiment, the test groups of cows were stoppedfeeding on the cysteamine-containing diet. However, the quantity of milkfat in the milk produced thereby was still higher when compared to thatof the control group of cows. This illustrates that the effect ofcysteamine lasted for some time after the administration of thecomposition was stopped. TABLE 21 Effect of the composition on 3.5 wt %FCM yield, kg/cow/day Group Time Test Group I Test Group II Test GroupIII Control Group Before 28.22 ± 2.37^(a) 29.81 ± 2.48^(ab) 28.35 ±3.99^(abc) 29.73 ± 3.35^(abcd) experiment State one 1^(st) to 20^(th)day 29.76 ± 2.10^(a) 27.44 ± 2.63^(B) 28.94 ± 2.28^(BC) 28.74 ±1.96^(BCD) 21^(st) to 37^(th) day 30.37 ± 2.55^(a) 30.93 ± 3.40^(ab)29.85 ± 3.17^(abc) 29.12 ± 2.28^(abcd) Average: 1^(st) to 37^(th) 30.07± 3.11^(a) 29.19 ± 3.30^(ab) 29.70 ± 2.17^(abc) 29.01 ± 2.17^(abcd) dayStage two 27.75 ± 2.30^(a) 26.72 ± 1.23^(ab) 26.37 ± 3.32^(abc) 25.41 ±1.42^(cd) Stage three 27.31 ± 1.25^(a) 29.08 ± 2.07^(ab)(Note:In the table, same letters in a row means that the difference of theirvalues is insignificant, i.e. p > 0.05, different letters in a same rowmeans that the difference of their values is significant p < 0.05;different letter size in a column means their values are significant,i.e. p < 0.01)

Table 21 above shows that the 3.5 wt % FCM yield of the Group I of cowsincreased by 3.55% (=[29.76−28.74]/28.74×100%) during 1^(st) to 20^(th)day of the experiment and by 4.29% (=[30.37−29.12]/29.12×100%) duringthe 21^(st) to 37^(th) day of the experiment as compared to that of thecontrol group of cows. This illustrates that the composition is alsoeffective in increasing the 3.5% FCM yield.

In stage two of the experiment, although the three test groups of cowswere stopped feeding on the cysteamine-containing diet, their 3.5 wt %FCM yield was higher than the control group of cows that was feeding onthe cysteamine-containing diet. This illustrates that the effect ofcysteamine persists for a while after administration thereof was stoppedand that it takes some time for cysteamine to take effect in thephysiology of the animals. TABLE 22 Effect of the composition on milkprotein content (wt %) Group Control Time Test Group I Test Group IITest Group III Group Before 2.63 ± 0.24^(a) 2.81 ± 0.18^(ab) 2.78 ±0.20^(abc) 2.67 ± experiment 0.29^(abcd) State one 1^(st) to 20^(th) day2.73 ± 0.21^(a) 2.85 ± 0.20^(ab) 2.85 ± 0.23^(abc) 2.81 ± 0.19^(abcd)21^(st) to 37^(th) day 2.70 ± 0.22^(a) 2.89 ± 0.14^(ab) 2.80 ±0.22^(abc) 2.83 ± 0.10^(abcd) Average: 1^(st) to 2.72 ± 0.21^(a) 2.87 ±0.17^(b) 2.83 ± 0.22^(abc) 2.82 ± 37^(th) day 0.14^(abcd) Stage two 2.65± 0.17^(a) 2.84 ± 0.09^(b) 2.72 ± 0.19^(ab) 2.76 ± 0.10^(ac) Stage three2.73 ± 0.18^(a) 2.89 ± 0.11^(b)(Note:In the table, same letters in a row means that the difference of theirvalues is insignificant, i.e. p > 0.05, different letters in a same rowmeans that the difference of their values is significant p < 0.05;different letter size in a column means their values are significant,i.e. p < 0.01)

TABLE 23 Effect of the composition on milk protein yield (g/cow/day)Group Time Test Group I Test Group II Test Group III Control GroupBefore 696.88 ± 63.64^(a) 754.59 ± 49.45^(ab) 744.93 ± 52.86^(abc)713.29 ± 76.97^(abcd) experiment State one 1^(st) to 20^(th) day 797.97± 61.72^(a) 766.37 ± 52.56^(ab) 757.05 ± 61.33^(abc) 789.33 ±52.52^(abcd) 21^(st) to 37^(th) day 731.09 ± 58.21^(a) 786.03 ±39.39^(b) 736.74 ± 56.89^(ac) 744.99 ± 25.83^(acd) Average: 1^(st) to764.53 ± 58.24^(a) 776.15 ± 45.16^(ab) 746.90 ± 59.58^(abc) 767.16 ±40.93^(abcd) 37^(th) day Stage two 714.85 ± 46.94^(a) 737.67 ±23.49^(ab) 692.29 ± 47.25^(ac) 691.23 ± 23.95^(acD) Stage three 696.00 ±45.47^(a) 781.20 ± 30.82^(b)(Note:In the table, same letters in a row means that the difference of theirvalues is insignificant, i.e. p > 0.05, different letters in a same rowmeans that the difference of their values is significant p < 0.05;different letter size in a column means their values are significant,i.e. p < 0.01)

Table 23 above similarly shows the effect of the composition althoughthe milk protein yield is expressed in gram per cow per day.

Table 23 shows that when the control group of cows was administered with40 g of cysteamine per cow every other day, the milk protein contentincreased by 5.86% (=[2.89 wt %−2.73 wt %]/2.73 wt %×100%). Thisillustrates that administering cysteamine at a relatively high doseevery other day can be more effective in increasing the milk proteinyield than administering cysteamine at a relatively low dose daily.TABLE 24 Number of somatic cells in milk produced by the cows (×10³/ml)Group Time Test Group I Test Group II Test Group III Control GroupBefore 1097.4 ± 1070.2^(a)   803 ± 1113.4^(ab)  126.9 ± 87.1^(abc)  99.9± 136.5^(abcd) experiment State one 1^(st) to 20^(th) day 296.25 ±452.33^(a) 727.63 ± 917.02^(ab) 395.50 ± 643.44^(abc) 129.63 ±152.56^(acd) 21^(st) to 37^(th) day 360.25 ± 479.59^(a) 613.38 ±556.70^(ab) 220.00 ± 348.65^(abc)  525.5 ± 917.59^(abcd) Average: 1^(st)to 328.25 ± 451.56^(a) 670.50 ± 735.22^(ab) 307.75 ± 508.08^(ac) 327.56± 667.51^(acd) 37^(th) day Stage two  322.9 ± 413.5^(a) 385.00 ±304.9^(ab)  187.1 ± 203.6^(abc)  224.4 ± 319.2^(abcd) Stage three 399.30± 560.40^(a) 335.50 ± 334.00^(ab)(Note:In the table, same letters in a row means that the difference of theirvalues is insignificant, i.e. p > 0.05, different letters in a same rowmeans that the difference of their values is significant p < 0.05;different letter size in a column means their values are significant,i.e. p < 0.01)

Table 24 above shows that, in the test Group I of cows, the number ofsomatic cells in milk decreased significantly. Similar decrease isobserved in the milk produced by the test Group II of cows. The numberof somatic cells in milk produced by control group of cows howeverincreased. Since the number of somatic cells in milk is generallyindicative of the overall health of the animals, the above dataillustrates that the overall health of dairy animals administered withcysteamine will improve.

The above experimental results illustrate that cysteamine or acysteamine composition is effective for improving lactation of lactatinganimals.

The contents of each of the references discussed above, including thereferences cited therein, are herein incorporated by reference in theirentirety. It is to be noted that numerous variations, modifications, andfurther embodiments are possible and accordingly, all such variations,modifications and embodiments are to be regarded as being within thescope of the present invention.

1. A method for improving lactation of lactating animals comprisingadministering to said animals an effective amount of cysteamine, a saltthereof or a composition containing cysteamine or a salt thereof.
 2. Themethod according to claim 1 wherein said improving lactation is anincrease in milk yields.
 3. The method according to claim 1 wherein saidimproving lactation is an increase in fat-corrected milk yields.
 4. Themethod according to claim 1 wherein said improving lactation is anincrease in mild fat content therein.
 5. The method according to claim 1wherein said improving lactation is an increase in milk protein contenttherein.
 6. The method according to claim 1 wherein said compositioncomprises substantially 1 to 95 wt % cysteamine having the chemicalformula of NH₂—CH₂—CH₂—SH, or a salt thereof.
 7. The method according toclaim 6 wherein said cysteamine-containing composition comprisessubstantially 30 wt % cysteamine.
 8. The method according to claim 1wherein said composition comprises substantially 1 to 80 wt % of astabilizer.
 9. The method according to claim 8 wherein said stabilizeris selected from the group consisting of cyclodextrin or a derivativethereof.
 10. The method according to claim 8 wherein said compositioncomprises substantially 10 wt % of said stabilizer.
 11. The methodaccording to claim 1 wherein said composition further comprisesingredient(s) selected from the group consisting of a bulking agent, adisintegration agent and a coated carrier.
 12. The method according toclaim 11 wherein said coated carrier is a solid carrier.
 13. The methodaccording to claim 11 wherein said coated carrier is has a coatingsoluble in intestines of said animals.
 14. The method according to claim11 wherein said coated carrier exhibits a multi-layer structure in saidcomposition.
 15. The method according to claim 11 wherein said coatedcarrier is adapted to remain un-dissolved at a pH of from about 1.5 to3.5.
 16. The method according to claim 1 wherein said lactating animalsare dairy cows.
 17. A feed for improving lactation of lactating animalscomprising a composition containing cysteamine or a salt thereof and astabilizer.
 18. The feed according to claim 17 wherein said improvinglactation is an increase in milk yields.
 19. The feed according to claim17 wherein said improving lactation is an increase in fat-corrected milkyields.
 20. The feed according to claim 17 wherein said improvinglactation is an increase in milk fat content therein.
 21. The feedaccording to claim 17 wherein said improving lactation is an increase inmild protein content therein.
 22. The feed according to claim 17 whereinsaid composition comprises substantially 1 to 95 wt % cysteamine havingthe chemical formula of NH₂—CH₂—CH₂—SH, or a salt thereof.
 23. The feedaccording to claim 22 wherein said cysteamine-containing compositioncomprises substantially 30 wt % cysteamine.
 24. The feed according toclaim 17 wherein said stabilizer is selected from the group consistingof cyclodextrin or a derivative thereof.
 25. The feed according to claim17 wherein said composition comprises substantially 1 to 80 wt % of saidstabilizer.
 26. The feed according to claim 17 wherein said compositionfurther comprises ingredient(s) selected from the group consisting of abulking agent, a disintegration agent and a coated carrier.
 27. The feedaccording to claim 26 wherein said coated carrier is a solid carrier.28. The feed according to claim 26 wherein said coated carrier has acoating soluble in intestines of said animals.
 29. The feed according toclaim 26 wherein said coated carrier exhibits a multi-layer structure insaid composition.
 30. The feed according to claim 26 wherein said coatedcarrier is adapted to remain undissolved at a pH of from about 1.5 to3.5.
 31. A The feed according to claim 17 wherein said lactating animalsare dairy cows.
 32. The feed according to claim 17 wherein the feedcomprises substantially 400 to 1000 ppm of said composition.
 33. Thefeed according to claim 17 wherein in its dry state, the feed comprisingcomprises substantially 1000 to 2174 ppm of said composition.
 34. Thefeed according to claim 17 wherein the feed comprises substantially 120to 300 ppm of cysteamine.
 35. The feed according to wherein the feed inits dry state comprises substantially 200 to 650 ppm of cysteamine. 36.The feed according to further comprising other foodstuffs selected fromthe group consisting of normal premix, cornmeal, cotton seed, wheatgluten, maize silage rutabaga, sugar beet pulp, apple pulp, ryegrass,fescue grass, alfalfa, feed concentrate and feed supplement.
 37. Amethod for improving lactation of lactating animals comprising: (a)producing a final feed by mixing a composition containing cysteamine ora salt thereof and a stabilizer with a suitable basal feed for saidanimals; and (b) feeding said animals with said final feed.
 38. Themethod according to claim 37 wherein said mixing in said step (a)comprises directly mixing said composition with said basal feed.
 39. Themethod according to claim 37 wherein said mixing in said step (a)comprises firstly preparing a premix including cysteamine or saidcomposition, and subsequently mixing said pre-mix with said basal feed.40. The method according to claim 39 wherein said premix is prepared bymixing cysteamine or said composition with a food material selected froma the group including consisting of cornmeal.
 41. The method accordingto claim 39 wherein said premix comprises 5 to 25 wt % of saidcomposition.
 42. The method according to claim 39 wherein said premixcomprises 10 to 20 wt % of said composition.
 43. The method according toclaim 37 further comprising feeding said animals with substantially 5.64to 12.71 g cysteamine, or a salt thereof, per animal per day.
 44. Themethod according to claim 37 further comprising feeding said animalswith substantially 18.79 to 42.36 of said composition per animal perday.